arXiv subjects

Hari Shroff

Publications and source records attributed to Hari Shroff.

13 recordsLinked to original sources

arXiv2025

DeepPD: Joint Phase and Object Estimation from Phase Diversity with Neural Calibration of a Deformable Mirror

Sample-induced aberrations and optical imperfections limit the resolution of fluorescence microscopy. Phase diversity is a powerful technique that leverages complementary phase information in sequentially acquired images with deliberately introduced aberrations--the phase diversities--to enable phase and object reconstruction and restore diffraction-limited resolution. These phase diversities are typically introduced into the optical path via a deformable mirror. Existing phase-diversity-based methods are limited to Zernike modes, require large numbers of diversity images, or depend on accurate mirror calibration--which are all suboptimal. We present DeepPD, a deep learning-based framework that combines neural representations of the object and wavefront with a learned model of the deformable mirror to jointly estimate both object and phase from only five images. DeepPD improves robustness and reconstruction quality over previous approaches, even under severe aberrations. We demonstrate its performance on calibration targets and biological samples, including immunolabeled myosin in fixed PtK2 cells.

physics.optics↗

arXiv2024

Convolutional Neural Network Transformer (CNNT) for Fluorescence Microscopy image Denoising with Improved Generalization and Fast Adaptation

Deep neural networks have been applied to improve the image quality of fluorescence microscopy imaging. Previous methods are based on convolutional neural networks (CNNs) which generally require more time-consuming training of separate models for each new imaging experiment, impairing the applicability and generalization. Once the model is trained (typically with tens to hundreds of image pairs) it can then be used to enhance new images that are like the training data. In this study, we proposed a novel imaging-transformer based model, Convolutional Neural Network Transformer (CNNT), to outperform the CNN networks for image denoising. In our scheme we have trained a single CNNT based backbone model from pairwise high-low SNR images for one type of fluorescence microscope (instance structured illumination, iSim). Fast adaption to new applications was achieved by fine-tuning the backbone on only 5-10 sample pairs per new experiment. Results show the CNNT backbone and fine-tuning scheme significantly reduces the training time and improves the image quality, outperformed training separate models using CNN approaches such as - RCAN and Noise2Fast. Here we show three examples of the efficacy of this approach on denoising wide-field, two-photon and confocal fluorescence data. In the confocal experiment, which is a 5 by 5 tiled acquisition, the fine-tuned CNNT model reduces the scan time form one hour to eight minutes, with improved quality.

q-bio.QM↗

arXiv2024

Prediction of Cellular Identities from Trajectory and Cell Fate Information

Determining cell identities in imaging sequences is an important yet challenging task. The conventional method for cell identification is via cell tracking, which is complex and can be time-consuming. In this study, we propose an innovative approach to cell identification during early $\textit{C. elegans}$ embryogenesis using machine learning. Cell identification during $\textit{C. elegans}$ embryogenesis would provide insights into neural development with implications for higher organisms including humans. We employed random forest, MLP, and LSTM models, and tested cell classification accuracy on 3D time-lapse confocal datasets spanning the first 4 hours of embryogenesis. By leveraging a small number of spatial-temporal features of individual cells, including cell trajectory and cell fate information, our models achieve an accuracy of over 91%, even with limited data. We also determine the most important feature contributions and can interpret these features in the context of biological knowledge. Our research demonstrates the success of predicting cell identities in time-lapse imaging sequences directly from simple spatio-temporal features.

q-bio.QM↗

arXiv2024

Live-cell imaging powered by computation

The proliferation of microscopy methods for live-cell imaging offers many new possibilities for users but can also be challenging to navigate. We focus here on computational methods that promise to boost live-cell fluorescence microscopy, where intra-cellular dynamics and cell viability constrain measurements considerably. Considering the tradeoffs between signal-to-noise ratio (SNR), spatial resolution, temporal resolution, and multi-color and multi-channel capacity, we review computational methods that can be layered on top of commonly used existing microscopies, as well as hybrid methods that integrate computation and microscope hardware.

q-bio.QM↗

arXiv2023

Phase Diverse Phase Retrieval for Microscopy: Comparison of Gaussian and Poisson Approaches

Phase diversity is a widefield aberration correction method that uses multiple images to estimate the phase aberration at the pupil plane of an imaging system by solving an optimization problem. This estimated aberration can then be used to deconvolve the aberrated image or to reacquire it with aberration corrections applied to a deformable mirror. The optimization problem for aberration estimation has been formulated for both Gaussian and Poisson noise models but the Poisson model has never been studied in microscopy nor compared with the Gaussian model. Here, the Gaussian- and Poisson-based estimation algorithms are implemented and compared for widefield microscopy in simulation. The Poisson algorithm is found to match or outperform the Gaussian algorithm in a variety of situations, and converges in a similar or decreased amount of time. The Gaussian algorithm does perform better in low-light regimes when image noise is dominated by additive Gaussian noise. The Poisson algorithm is also found to be more robust to the effects of spatially variant aberration and phase noise. Finally, the relative advantages of re-acquisition with aberration correction and deconvolution with aberrated point spread functions are compared.

eess.IV↗

arXiv2022

A Semi-automatic Cell Tracking Process Towards Completing the 4D Atlas of C. elegans Development

The nematode Caenorhabditis elegans (C. elegans) is used as a model organism to better understand developmental biology and neurobiology. C. elegans features an invariant cell lineage, which has been catalogued and observed using fluorescence microscopy images. However, established methods to track cells in late-stage development fail to generalize once sporadic muscular twitching has begun. We build upon methodology which uses skin cells as fiducial markers to carry out cell tracking despite random twitching. In particular, we present a cell nucleus segmentation and tracking procedure which was integrated into a 3D rendering GUI to improve efficiency in tracking cells across late-stage development. Results on images depicting aforementioned muscle cell nuclei across three test embryos suggest the fiducial markers in conjunction with a classic tracking paradigm overcome sporadic twitching.

cs.CV↗

arXiv2022

Coiling of cellular protrusions around extracellular fibers

Protrusions at the leading-edge of a cell play an important role in sensing the extracellular cues, during cellular spreading and motility. Recent studies provided indications that these protrusions wrap (coil) around the extra-cellular fibers. The details of this coiling process, and the mechanisms that drive it, are not well understood. We present a combined theoretical and experimental study of the coiling of cellular protrusions on fibers of different geometry. Our theoretical model describes membrane protrusions that are produced by curved membrane proteins that recruit the protrusive forces of actin polymerization, and identifies the role of bending and adhesion energies in orienting the leading-edges of the protrusions along the azimuthal (coiling) direction. Our model predicts that the cell's leading-edge coils on round fibers, but the coiling ceases for a fiber of elliptical (flat) cross-section. These predictions are verified by 3D visualization and quantitation of coiling on suspended fibers using Dual-View light-sheet microscopy (diSPIM). Overall, we provide a theoretical framework supported by high spatiotemporal resolution experiments capable of resolving coiling of cellular protrusions around extracellular fibers of varying diameters.

physics.bio-ph↗

arXiv2021

Multiple Hypothesis Hypergraph Tracking for Posture Identification in Embryonic Caenorhabditis elegans

Current methods in multiple object tracking (MOT) rely on independent object trajectories undergoing predictable motion to effectively track large numbers of objects. Adversarial conditions such as volatile object motion and imperfect detections create a challenging tracking landscape in which established methods may yield inadequate results. Multiple hypothesis hypergraph tracking (MHHT) is developed to perform MOT among interdependent objects amid noisy detections. The method extends traditional multiple hypothesis tracking (MHT) via hypergraphs to model correlated object motion, allowing for robust tracking in challenging scenarios. MHHT is applied to perform seam cell tracking during late-stage embryogenesis in embryonic C. elegans.

eess.IV↗

arXiv2021

An Exact Hypergraph Matching Algorithm for Nuclear Identification in Embryonic Caenorhabditis elegans

Finding an optimal correspondence between point sets is a common task in computer vision. Existing techniques assume relatively simple relationships among points and do not guarantee an optimal match. We introduce an algorithm capable of exactly solving point set matching by modeling the task as hypergraph matching. The algorithm extends the classical branch and bound paradigm to select and aggregate vertices under a proposed decomposition of the multilinear objective function. The methodology is motivated by Caenorhabditis elegans, a model organism used frequently in developmental biology and neurobiology. The embryonic C. elegans contains seam cells that can act as fiducial markers allowing the identification of other nuclei during embryo development. The proposed algorithm identifies seam cells more accurately than established point-set matching methods, while providing a framework to approach other similarly complex point set matching tasks.

cs.CV↗

arXiv2020

Spatio-angular fluorescence microscopy III. Constrained angular diffusion, polarized excitation, and high-NA imaging

We investigate rotational diffusion of fluorescent molecules in angular potential wells, the excitation and subsequent emissions from these diffusing molecules, and the imaging of these emissions with high-NA aplanatic optical microscopes. Although dipole emissions only transmit six low-frequency angular components, we show that angular structured illumination can alias higher frequency angular components into the passband of the imaging system. We show that the number of measurable angular components is limited by the relationships between three time scales: the rotational diffusion time, the fluorescence decay time, and the acquisition time. We demonstrate our model by simulating a numerical phantom in the limits of fast angular diffusion, slow angular diffusion, and weak potentials.

physics.optics↗

arXiv2019

Spatio-angular fluorescence microscopy II. Paraxial 4$f$ imaging

We investigate the properties of a single-view fluorescence microscope in a 4$f$ geometry when imaging fluorescent dipoles without using the monopole or scalar approximations. We show that this imaging system has a spatio-angular band limit, and we exploit the band limit to perform efficient simulations. Notably, we show that information about the out-of-plane orientation of ensembles of in-focus fluorophores is recorded by paraxial fluorescence microscopes. Additionally, we show that the monopole approximation may cause biased estimates of fluorophore concentrations, but these biases are small when the sample contains either many randomly oriented fluorophores in each resolvable volume or unconstrained rotating fluorophores.

physics.optics↗

arXiv2019

Spatio-angular fluorescence microscopy I. Basic theory

We introduce the basic elements of a spatio-angular theory of fluorescence microscopy, providing a unified framework for analyzing systems that image single fluorescent dipoles and ensembles of overlapping dipoles that label biological molecules. We model an aplanatic microscope imaging an ensemble of in-focus fluorescent dipoles as a linear Hilbert-space operator, and we show that the operator takes a particularly convenient form when expressed in a basis of complex exponentials and spherical harmonics$-$a form we call the dipole spatio-angular transfer function. We discuss the implications of our analysis for all quantitative fluorescence microscopy studies and lay out a path towards a complete theory.

physics.optics↗

arXiv2009

Mechanical conversion of low-affinity Integration Host Factor binding sites into high-affinity sites

Although DNA is often bent in vivo, it is unclear how DNA-bending forces modulate DNA-protein binding affinity. Here, we report how a range of DNA-bending forces modulates the binding of the Integration Host Factor (IHF) protein to various DNAs. Using solution fluorimetry and electrophoretic mobility shift assays, we measured the affinity of IHF for DNAs with different bending forces and sequence mutations. Bending force was adjusted by varying the fraction of double-stranded DNA in a circular substrate, or by changing the overall size of the circle (1). DNA constructs contained a pair of Forster Resonance Energy Transfer dyes that served as probes for affinity assays, and read out bending forces measured by optical force sensors (2). Small bending forces significantly increased binding affinity; this effect saturated beyond ~3 pN. Surprisingly, when DNA sequences that bound IHF only weakly were mechanically bent by circularization, they bound IHF more tightly than the linear "high-affinity" binding sequence. These findings demonstrate that small bending forces can greatly augment binding at sites that deviate from a protein's consensus binding sequence. Since cellular DNA is subject to mechanical deformation and condensation, affinities of architectural proteins determined in vitro using short linear DNAs may not reflect in vivo affinities.

q-bio.BM↗